title: In vitro phenotypes to elvitegravir and dolutegravir in primary macrophages and lymphocytes of clonal recombinant viral variants selected in patients failing raltegravir.
abstract: OBJECTIVE
  The cross-resistance profiles of elvitegravir and dolutegravir on raltegravir-resistant variants is still controversial or not available in macrophages and lack extensive evaluations on wide panels of clonal variants. Thus, a complete evaluation in parallel with all currently available integrase inhibitors (INIs) was performed.
  
  
  METHODS
  The integrase coding region was RT-PCR-amplified from patient-derived plasma samples and cloned into an HIV-1 molecular clone lacking the integrase region. Twenty recombinant viruses bearing mutations to all primary pathways of resistance to raltegravir were phenotypically evaluated with each integrase inhibitor in freshly purified CD4+ T cells or monocyte-derived macrophages.
  
  
  RESULTS
  Y143R single mutants conferred a higher level of raltegravir resistance in macrophages [fold change (FC) 47.7-60.24] compared with CD4+ T cells (FC 9.55-11.56). All other combinations had similar effects on viral susceptibility to raltegravir in both cell types. Elvitegravir displayed a similar behaviour both in lymphocytes and macrophages with all the tested patterns. When compared with raltegravir, none to modest increases in resistance were observed for the Y143R/C pathways. Dolutegravir maintained its activity and cross-resistance profile in macrophages. Only Q148H/R variants had a reduced level of susceptibility (FC 5.48-18.64). No variations were observed for the Y143R/C (+/-T97A) or N155H variants.
  
  
  CONCLUSIONS
  All INIs showed comparable antiretroviral activity in both cell types even if single mutations were associated with a different level of susceptibility in vitro to raltegravir and elvitegravir in macrophages. In particular, dolutegravir was capable of inhibiting with similar potency infection of raltegravir-resistant variants with Y143 or N155 pathways in both HIV-1 major cell reservoirs.
authors.affiliation: Department of Clinical and Experimental Medicine, Università degli Studi dell'Insubria, Varese, Italy.
authors.forename: Filippo
authors.initials: F
authors.lastname: Canducci
authors.forename: Elisa Rita
authors.initials: ER
authors.lastname: Ceresola
authors.forename: Diego
authors.initials: D
authors.lastname: Saita
authors.forename: Antonella
authors.initials: A
authors.lastname: Castagna
authors.forename: Nicola
authors.initials: N
authors.lastname: Gianotti
authors.forename: Mark
authors.initials: M
authors.lastname: Underwood
authors.forename: Roberto
authors.initials: R
authors.lastname: Burioni
authors.forename: Adriano
authors.initials: A
authors.lastname: Lazzarin
authors.forename: Massimo
authors.initials: M
authors.lastname: Clementi
mesh_terms: D019380:Anti-HIV Agents / Q000494:pharmacology* / Q000627:therapeutic use; D002478:Cells, Cultured; D003001:Cloning, Molecular; D024882:Drug Resistance, Viral; D015658:HIV Infections / Q000188:drug therapy / Q000821:virology; D019427:HIV Integrase / Q000235:genetics; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification; D006575:Heterocyclic Compounds, 3-Ring / Q000494:pharmacology* / Q000627:therapeutic use; D006801:Humans; D008214:Lymphocytes / Q000821:virology*; D008264:Macrophages / Q000821:virology*; D010078:Oxazines; D010879:Piperazines; D011728:Pyridones; D011760:Pyrrolidinones / Q000494:pharmacology* / Q000627:therapeutic use; D015363:Quinolones / Q000494:pharmacology* / Q000627:therapeutic use; D012367:RNA, Viral / Q000235:genetics; D000068898:Raltegravir Potassium; D011995:Recombination, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D000099226:Dolutegravir
chemical_list: D019380:Anti-HIV Agents; D019427:HIV Integrase; D006575:Heterocyclic Compounds, 3-Ring; D010078:Oxazines; D010879:Piperazines; D011728:Pyridones; D011760:Pyrrolidinones; D015363:Quinolones; D012367:RNA, Viral; D000068898:Raltegravir Potassium; D000099226:Dolutegravir; C509700:elvitegravir
keywords: HIV eradication; HIV reservoir; HIV therapy; cross-resistance; viral fitness